Selection of Aptamers with Large Hydrophobic 2′-Substituents,Journal of the American Chemical Society

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Selection of Aptamers with Large Hydrophobic 2′-Substituents
Journal of the American Chemical Society ( IF 14.4 ) Pub Date : 2020-01-21 , DOI: 10.1021/jacs.9b10538
Qian Shao ₁ , Tingjian Chen ₁ , Kai Sheng ₁ , Zhixia Liu ₁ , Zhuochen Zhang ₁ , Floyd E Romesberg ₁

Affiliation

Previously, we evolved a DNA polymerase, SFM4-3, for the recognition of substrates modified at their 2' positions with a fluoro, O-methyl, or azido substituent. Here we use SFM4-3 to synthesize 2'-azido-modified DNA; we then use the azido group to attach different, large hydrophobic groups via click chemistry. We show that SFM4-3 recognizes the modified templates under standard conditions, producing natural DNA and thereby allowing amplification. To demonstrate the utility of this remarkable property, we use SFM4-3 to select aptamers with large hydrophobic 2' substituents that bind human neutrophil elastase or the blood coagulation protein factor IXa. The results indicate that SFM4-3 should facilitate the discovery of aptamers that adopt novel and perhaps more protein-like folds with hydrophobic cores that in turn allow them to access novel activities.

中文翻译：

具有大疏水性 2'-取代基的适体的选择

以前，我们开发了一种 DNA 聚合酶 SFM4-3，用于识别在其 2' 位置用氟、O-甲基或叠氮基取代基修饰的底物。这里我们使用 SFM4-3 来合成 2'-叠氮基修饰的 DNA；然后我们使用叠氮基通过点击化学连接不同的大疏水基团。我们表明 SFM4-3 在标准条件下识别修改后的模板，产生天然 DNA，从而允许扩增。为了证明这一非凡特性的效用，我们使用 SFM4-3 来选择具有大疏水性 2' 取代基的适体，这些取代基与人类中性粒细胞弹性蛋白酶或凝血蛋白因子 IXa 结合。

更新日期：2020-01-21

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